1. AβOs induce hypothalamic inflammation, eIF2α phosphorylation and impaired insulin signaling A, BWestern blot analysis of eIF2α‐P levels in the hypothalamus of mice 4 h (A; n = 4 animals/group) or 7 days (B; n = 6 Veh; 5 AβOs) after a single i.c.v. injection of vehicle (Veh) or 10 pmol AβOs.
AβOs induce hypothalamic inflammation, eIF2α phosphorylation and impaired insulin signaling A, BWestern blot analysis of eIF2α‐P levels in the hypothalamus of mice 4 h (A; n = 4 animals/group) or 7 days (B; n = 6 Veh; 5 AβOs) after a single i.c.v. injection of vehicle (Veh) or 10 pmol AβOs. Graphs show densitometric data normalized by total eIF2α levels. *P =  CWestern blot analysis of ATF4 levels in the hypothalamus of mice 4 h after i.c.v. injection of vehicle (Veh) or 10 pmol AβOs; graph shows densitometric data normalized by β‐actin (n = 7 Veh; 8 AβOs). #P = 0.0731; Student's t‐test. D, EWestern blot analysis of hypothalamic phospho‐IKKβ levels in the hypothalamus of mice 4 h (C; n = 6 animals/group) or 7 days (D; n = 4 Veh; 5 AβOs) after i.c.v. injection of vehicle or 10 pmol AβOs. Graphs show densitometric data normalized by total IKKβ levels. In (D), *P = 0.0437; in (E), *P = 0.0444; Student's t‐test. F, GWestern blot analysis of IκBα (F; n = 6 animals/group) and cytoplasmic phospho‐p65‐NF‐κB (G; n = 4 Veh; 5 AβOs) in the hypothalamus of mice 4 h after i.c.v. injection of vehicle or 10 pmol AβOs. Graphs show densitometric data normalized by actin (F) or total NF‐κB levels (G). *P =  HNuclear NF‐κB levels in the hypothalamus 6 h after i.c.v. injection of vehicle or 10 pmol AβOs in mice. Graphs show NF‐κB levels normalized by nuclear marker, lamin B (n = 6 animals/group). **P = 0.0024; Student's t‐test. I, JIRS‐1pSer636 (I; n = 4 animals/group) and pTyr465 (J; n = 6 Veh; 4 AβOs) levels in the hypothalamus 7 days after i.c.v. injection of vehicle or AβOs in mice. Graphs show IRS‐1pSer or IRS‐1pTyr levels normalized by total IRS‐1. In (I), *P = 0.0043; in (J), *P = 0.0275; Student's t‐test. KTwelve‐hour food intake after a single i.c.v. infusion of insulin (200 mU) in mice. Experiment was performed 7 days after i.c.v. injection of vehicle or AβOs (n = 5 PBS; 5 Veh + Insulin; 9 AβOs + Insulin), data are representative of two independent experiments with similar results. ***P < 0.0001, one‐way ANOVA followed by Bonferroni post hoc test comparing Veh‐Insulin versus PBS groups. Data information: Data are expressed as means ± SEM. In (A–J), to assess statistical significance, AβO‐injected mice were compared to vehicle‐injected mice.Source data are available online for this figure.
Julia R Clarke et al. EMBO Mol Med. 2015;7:
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